In vitro migration of murine MDSCs was evaluated in 24-well plates with transwell polycarbonate-permeable supports (8.0 μm; Costar Corning, Cambridge, MA, USA). MDSCs (5 × 105; >80% purity) were plated in the upper chambers of the inserts 30 min after incubation with a CXCR2 antagonist at each concentration, and recombinant murine CXCL1, CXCL2, and CXCL5 (PeproTech) were placed in the lower chamber at a concentration of 1, 10, or 100 ng/mL. After incubation for 3 h, the number of MDSCs in the bottom compartment was counted using Acubright counting beads (Life Technologies, Carlsbad, CA, USA)17 (link).
To evaluate the migration of human MDSCs, MDSCs (1 × 105; >80% purity) were plated in the upper chambers of the inserts 30 min after incubation with the CXCR2 antagonist at each concentration, and recombinant human CXCL1, CXCL2, and CXCL5 (PeproTech) were placed in the lower chamber at a concentration of 1 or 10 ng/ml. After incubation for 6 h, the number of MDSCs in the bottom compartment was counted36 (link).
To evaluate the migration of human MDSCs, MDSCs (1 × 105; >80% purity) were plated in the upper chambers of the inserts 30 min after incubation with the CXCR2 antagonist at each concentration, and recombinant human CXCL1, CXCL2, and CXCL5 (PeproTech) were placed in the lower chamber at a concentration of 1 or 10 ng/ml. After incubation for 6 h, the number of MDSCs in the bottom compartment was counted36 (link).
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